CIHM 
Microfiche 
Series 
({Monographs) 


ICIMH 

Collection  de 
microfiches 
(monographies) 


Canadian  Institute  for  Historical  Microraproductions  /  Institut  Canadian  da  microraproductions  historiquas 


Technical  and  Bibliographic  Notes  /  Notes  techniques  et  bibliographiques 


The  Institute  has  attempted  to  obtain  the  best  original 
copy  available  for  filming.  Features  of  this  copy  which 
may  be  bibliograpliically  unique,  which  may  alter  any  of 
the  images  in  the  reproduction,  or  which  may 
significantly  change  the  usual  method  of  filming  are 
checked  below. 


D 
D 


n 
n 


D 


Coloured  covers  / 
Couverture  de  couleur 

Covers  damaged  / 
Couverture  endommag^e 

Covers  restored  and/or  laminated  / 
Couverture  restaur^e  et/ou  peiiicul^e 


^ 


Cover  title  missing  /  Le  titre  de  couverture  manque 

Coloured  maps  /  Cartes  g^ographiques  en  couleur 

Coloured  ink  (i.e.  other  than  blue  or  black)  / 
Encre  de  couleur  (i.e.  autre  que  bieue  ou  noire) 

Coloured  plates  and/or  illustrations  / 
Planches  et/ou  illustrations  en  couleur 

Bound  with  other  material  / 
Relid  avec  d'autres  documents 

Only  edition  available  / 
Seule  Edition  disponible 

Tight  binding  may  cause  shadows  or  distortion  along 
interior  margin  /  La  reliure  serr^e  peut  causer  de 
I'ombre  ou  de  la  distorsion  le  long  de  la  marge 
int^rieure. 

Blank  leaves  added  during  restorations  may  appear 
within  the  text.  Whenever  possible,  these  have  been 
omitted  from  filming  /  II  se  peut  que  certaines  pages 
blanches  ajout^es  lors  d'une  restauration 
apparaissent  dans  le  texte,  mais,  lorsque  cela  4tait 
possible,  ces  pages  n'ont  pas  ixi  fiimees. 


L'Institut  a  microfilm^  le  meilleur  exemplaire  qu'il  lul  a 
6\6  possible  de  se  procurer.  Les  details  de  cet  exem- 
plaire qui  son!  peut-£tre  uniques  du  point  de  vue  bibli- 
ographique,  qui  peuvent  modifier  une  image  reproduite, 
ou  qui  peuvent  exiger  une  modifk:ation  dans  la  m^tho- 
de  normale  de  filmage  sont  indiqu^s  cl-dessous. 

I     I  Coloured  pages  /  Pages  de  couleur 

I I  Pages  damaged/ Pages  endommag6es 

□  Pages  restored  and/or  laminated  / 
Pages  restaur^es  et/ou  pellicul^es 

0  Pages  discoloured,  stained  or  foxed  / 
Pages  ddcolordes,  tachet^es  ou  piqu^es 

I     I  Pages  detached  /  Pages  d6tach6es 

p^l  Showthrough / Transparence 

□  Quality  of  print  varies  / 
Quality  in^gale  de  I'impression 

Includes  supplementary  material  / 
Comprend  du  materiel  suppl^mentaire 

Pages  wholly  or  partially  obscured  by  errata  slips, 
tissues,  etc.,  have  been  refilmed  to  ensure  the  best 
possible  image  /  Les  pages  totalement  ou 
partiellement  obscurcies  par  un  feuillet  d'errata,  une 
pelure,  etc.,  ont  6t6  film6es  k  nouveau  de  fa^on  k 
obtenir  la  meilleure  image  possible. 

Opposing  pages  with  varying  colouration  or 
discolourations  are  filmed  twice  to  ensure  the  best 
possible  image  /  Les  pages  s'opposant  ayant  des 
colorations  variables  ou  des  decolorations  sont 
film6es  deux  fois  afin  d'obtenir  la  meilleure  image 
possible. 


D 
D 


D 


Additional  comments  / 
Commentaires  suppl^mentaires: 


Pagination  is  as  folloMs:     p.  23-%9. 


i         : 

Si 

This  item  Is  filmed  at  the  reduction  ratio  checked  below/ 

Ce  document  est  filme  au  taux  de  reduction  indiqui  ci-destous. 

lOx                             14x                            18x 

22x 

26x 

30x 

7 

1 
1 

12x 

16x 

20x 

24x 

28x 

32x 

Th«  copy  filmed  h«r«  has  b««n  raproduead  thanks 
to  tha  ganarosity  of: 

Blacker-Wood  Library  of  Biology 
HcClli  University,  Montreal 

Tha  imagas  appaaring  hara  ara  tha  baat  quality 
possibia  eonsidaring  tha  condition  and  lagibility 
of  tha  original  copy  and  in  kaaping  with  tha 
filming  contract  spacificationa. 


Original  copias  in  printad  papar  eovara  ara  fllmad 
baginning  with  tha  front  covar  and  anding  on 
tha  last  paga  with  a  printad  or  illustratad  impraa- 
sion,  or  tha  back  covar  whan  appropriata.  All 
othar  original  copiaa  ara  filmad  baginning  on  tha 
first  paga  with  a  printad  or  illustratad  impraa- 
sion,  and  anding  on  tha  laat  paga  with  a  printad 
or  illustratad  impraaaion. 


Tha  last  racordad  frama  on  aach  microflcha 
shall  contain  tha  symbol  ^-^  (maaning  "CON- 
TINUED"), or  tha  symbol  V  (maaning  "END"), 
whiehavar  appiias. 

Maps,  platas.  charts,  ate.  may  ba  filmad  at 
diffarant  raduction  ratios.  Thosa  too  larga  to  ba 
antiraly  includad  in  ona  axposura  an  filmad 
baginning  in  tha  uppar  laft  hand  cornar.  laft  to 
right  and  top  to  bonom,  as  many  framas  as 
raquirad.  Tha  following  diagrams  illustrata  tha 
mathod: 


L'axamplaira  filmi  fut  raproduit  grica  k  la 
gintrosit*  da: 

Blacker-Wood  Library  of  Biology 
McCin  University,  Montreal 


Las  imagas  suivsntas  ont  M  raproduitas  avac  la 
plus  grand  soin.  compta  tanu  da  la  condition  at 
da  la  nattat*  da  I'axampiaira  film*,  at  an 
conformity  avac  las  conditions  du  contrat  da 
filmaga. 

Las  axamplairas  originaux  dont  la  couvartura  an 
papiar  aat  ImprimAa  sont  film«s  an  commandant 
par  la  pramiar  plat  at  an  tarminant  soit  par  la 
darniira  paga  qui  comporta  una  amprainta 
d'imprassion  ou  d'illustration.  soit  par  la  sacond 
plat,  salon  la  cas.  Tous  las  autras  axamplairas 
originaux  sont  filmis  an  commandant  par  la 
pramiAra  paga  qui  comporta  una  amprainta 
d'impraasion  ou  d'illustration  at  an  tarminant  par 
la  darniira  paga  qui  comporta  una  talia 
amprainta. 

Un  das  symbolas  suivants  spparaitra  sur  la 
darnlAra  imaga  da  chaqua  microfiche,  salon  la 
cas:  la  symbols  — ^  signifia  "A  SUIVRE".  la 
symbols  ▼  signifia  "FIN". 

Las  cartas,  planchas.  tableaux,  etc..  peuvent  itre 
film«s  A  des  taux  da  reduction  diff«rents. 
Lorsque  la  document  est  trop  grand  pour  itra 
reproduit  en  un  seul  clich*.  il  est  film«  A  partir 
da  I'angia  sup«rieur  gauche,  de  gauche  A  droite, 
et  de  haut  an  bas.  en  prenant  la  nombra 
d'imagas  nAcessaira.  Las  diagrammes  suivants 
illustrent  la  mAthoda. 


1 

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MICIOCOPV   lESOlUTION   TiST  CHART 

(ANSI  and  ISO  TEST  CHART  No.  2) 


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^  /APPLIED  IIVMGE    Inc 

^^  1653   East    Main    Street 

S^S  Rochester.    New    Tork         U609        USA 

I^S  (ne)   *82  -  0300  -  Phone 

^aS  (716)    28S  -  59S9  ~  Fa» 


•.I   I    2^'MLt 


I  UNIV2S5??y 

'  ^  mju. 


CS  ON  MOLLUSC  AN  CELOMIC  FLUID     . 


F    CHANGE    IN    ENVIUONMENT    ON    THE 
GWaBON  DIOXIDE  CONTENT  OF  THE 
CELOMIC  FLUID 


ANAEROBIC  RESPIRATION  IN  MYA  ARENARIA 


BT 

J.   B.  COLLI  P 


(Fhom  thb  Marine  Biolooical  Statio.v,  Dui-AuruKt:  Bay,  Canada) 


.      i 


RlFBINTHD  FBOU 

THE  JOURNAL  OF  BIOLOGICAL  CHEMISTRY 
Vol.  XLV,  No.  1,  December,  1920 


INprihtril  fn.iii  I  in    loi  iiwi.  m  Hioi.oi.ical  ('tit.MJ-i  in,  \..l    MS  .  N.. 

IhllTlllKT,   I<I20 


STUDIES  ON  MOLLUSCAN  CELOMIC  FLUID. 

EFFECT   OF   CHANGE    IN   ENVIRONMENT    ON   THE    CARBON 
DIOXIDE  CONTENT  OF  THE  CELOMIC  FLUID. 

ANAEROBIC  RESPIRATION  IN  MYA  ARENARIA. 

Hy  .1.  \i.  COM.Il'. 

(From    the   Marine    liiologicnl   Stntinn.    h(i„irhii,-   li,,;,,    Ciinivlit.) 

(Rccoivcd  for  public:iticm,  Si-pUrriber  10.  ISWI.) 

INTHODICTIOV. 

It  was  noted  in  a  previous  coimininicalidii  fh  tliat  tlic  con- 
font  of  conit)inc(l  carlion  dioxide  of  molhisean  eclomic  flui<l  tends 
to  ri.se  when  the  animals  are  removed  from  llieir  natiiralVin  iron- 
ment  whereas  a  fall  was  noticed  in  this  factor  in  the  case  of  fisii 
removed  from  their  natural  hal.itat.  In  order  to  determine  what 
was  the  cause  of  this  peculiar  effect  in  the  molhisean  forms  a 
series  of  experiments  was  undertaken,  the  results  of  wliicii  are 
herein  reported. 

KXl'KUIMK.VTAL. 

EjJcd  of  Exposure  to  Atmoxphiric  Air  on  (lie  Comhinnl  Crhon 
Dioxide  Contcitl  of  the  C domic  Fluid. 

The  metliod  of  securinK  samples  of  celomic  fluid  or  "clam 
juice"  from  the  various  .specimens  was  the  .same  as  that  detailed 
previously  (1).  Specimens  of  seven  species  of  pelecyixxl  Mol- 
hisca  were  exposed  to  atmospheric  air  in  a  closed  jtlass  container 
for  varying  periods  of  time.  One  species  of  the  Ani|.hineura 
and  two  si)ecies  of  the  C.asfropoda  were  similarly  studied.  .Sev- 
eral non-mollu.scan  forms  were  al.so  exjiosed  to  ;itmospheric  air 
under  similar  con<Iitions.  These  inchu'ed  the  calcareous  shelled 
arthropod  Bulnnu.s  ufjuilh,  the  common  hrachiopod  Tinhrehlla 
Iranmraa,  vaiious  ("nistacea  of  the  decapod  typv,  starfish,  sea 
urchins,   .and   certain   varieties   of   marine   fish.     The    container 

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28 


Mollusejui  CVlnniic  F1ui<l 


used  wiis  of  nood  size  and  a  full  supi)ly  nf  oxyncii  was  assured. 
It  vas  k('i)t  covorod  to  i)r('V»'iit  Iciss  of  water  li.v  evaiioration. 
Filter  paper  nioistene<l  in  sea  water  was  fre<iueiitly  placed  in  the 
container  with  the  specimens.  Specimens  which  were  exposed 
to  air  were  kept  in  certain  instances  at  a  fairly  constant  tempera- 
ture l)y  iiumersiiijj  the  coiitainer  in  sea  water  while  in  others 
they  were  kept  in  the  laboratory  and  were  thus  subject  to  the 
temperature  changes  of  the  latter.  Table  I  illustrates  the  efTects 
of  ex)»osure  to  air  for  various  j)eri(»<ls  upon  the  combined  carbon 
(lioxi<le  content  of  the  celomic  fluid  of  the  different  species  inves- 
tigated. Tile  ra))id  increase  in  the  carbon  dioxide  content  of  the 
celomic  fluid  of  the  molluscan  forms  and  the  arthropol  Bahniis 
(ifiuilla  is  very  striking.  That  this  increa.se  is  due  to  bicarbonate 
is  evident  .since  the  samples  wen-  e(|uilibrated  with  atmospheric 
air  l)efore  being  submitted  to  analysis.  The  decapod  cnistaoeans 
examined  failed  to  show  this  reaction,  while  a  very  slight  increase 
was  in  some  instances  manifeste<l  in  the  echinoderms.  As  the 
latter  reaction  was  not  uniform  it  is  of  very  doubtful  significance. 
The  sunival  time  for  specimens  exposed  to  atmospheric  air 
varied  greatly.  It  was  early  noted  that  .!///«  nnuaria  was  pecu- 
liarly resistant  to  long  exposure  toatmosplu-ric  air  at  the  tempera- 
tures which  prevailed  in  the  surface  wat(M-  and  the  air  at  Depar- 
ture Bay  during  the  summer  months.  It  was  for  this  reason 
used  extt'nsively  in  lat<.>r  investigation.  It  is  regretted  that  no 
facilities  were  availal)lc  which  would  enable  one  to  keep  si)ecimcns 
at  a  low  as  well  as  constant  temperature.  The  results  obtained 
will  therefore  have  to  be  considered  in  the  light  of  this  condition. 
The  greatest  increase  in  the  carbon  dioxide  content  of  the  blood 
was  in  two  sjiecimens  of  Mija  airnaria  which  had  l)een  exposed 
f<ir  !)()  hours.  The  increa.se  here  was  from  ().5  volumes  per  cent 
in  the  controls  kept  in  sea  water  to  105  volumes  p«-r  cent  in  the 
specimens  cxpo.sed  to  atmospheric  air  a(  the  temperature  of  sur- 
face sea  water.  The  container  used  was  a  (1  liter  cylindrical 
glass  mu.seum  jar  and  it  was  opened  daily  both  for  the  pm-pose 
of  removing  specimens  for  examination  and  to  allow  a  change  of 
air.  The  temperatme  of  the  sea  water  in  the  vessel  which  was 
taken  on  the  4  consecutive  days  during  which  these  specimens 
wen;  exposed  was  li».S%  I8.S",  lit.:r,  and  1S.«I"('.     Slightlv  over  a 


♦*!*■:  ',. 


J.  B.  CoUip 


29 


sixlccnfold  im-rease  in  the  coniluiied  caibou  dioxide  in  the  blood 
was  noticed  in  this  instance. 

The  incmiso  in   the  caiLon  dioxide  in  the  small  pelecypod 
Macoma  secta  from  11.2  to  48.()  volumes  per  cent  in  6  hours  is 
noteworthy,  as  is  also  that  observed  in  the  gastropod  Polynices 
lewisii    following    80    hours    exposun".     The    combined    carbon 
dio.\ide  rose  in  tliis  latter  instance  from  12.5  volumes  per  cent 
in  the  control  to  77.4  volumes  per  cent  in  (he  ex|)o.sed  si)ecimen. 
There  were  few  forms  wliidi  would  surA  ive  an  eximsure  to  atmos- 
pheric air  of  more  than  24  hours  at  the  i)revailinK  land  and  sur- 
face^ water  temperatures.     The  amphineuran   Vniptochiton,    the 
cockle  Vnnlium  corbis.  and  the  eaple  barnacle   BalnnuH  aqnilla 
were  \-ery  .s>nsitiv(>  (o  exposure.     'J'he  horse  ••lam  Schiznlhoerm 
niilMli,  and  the  butler  clam  So.ridonius  ijiqmdea  withstood  an 
exposure  of  24  to  48  hours.     The  lit  ( le  neck  clam  Paphia  sUnnincn 
and  the  edible  form  Mija  tircnnrin  were  very  resistant  to  exposure 
and  in  some  instances  sur\iv(Hl  as  long  as  T)  days  when  placed 
in  the  air,  the  evaporation  of  water  beinj;  practically  excliuled. 
The  nudibranch  Anisoihris  was  most  .sensitive  of  all  forms,  dying 
shortly  after  being  brought   into  the  lal)oratory.     The  absence 
of  a  calcareous  shell  is  probably  a.ssociated  with  the  lack  of  resis- 
tance to  exposure  to  air  in  this  f(.rm  although  the  temperature 
factor  must  al.so  be  of  great  importjmce. 


Kfffd  of  Kxpimm'  (o  Mii,„xi>lin!c  Mr  on   tin:   Carhun  Dioxide 
Ciiltncilij  of  (he  Cdoniir  Flin'il. 

Several  samples  of  celomic  tluid  taken  from  specimens  exjwsed 
to  atmospheric  air  for  varying  periods  weiv  analyzed  in  the  Van 
Slyke  apparatus  (2)  when  eciuiiibrated  with  atmos[)heric  air  and 
also  when  equilibrated  with  alveolar  air  of  lli(<  normal  sui)ject 
after  the  manner  described  by  Van  Slyke  and  ('ullen  (;{).  Table 
II  illustrates  the  results  obtained  in  this  .series  of  exp(>riinents. 
It  will  be  noted  that  in  every  instance  the  carbon  dioxide  cai)acity 
of  the  sample  was  considerably  in  excess  of  the  carbon  dioxide 
content  of  the  same  when  equilibrated  with  atmospheric  air. 


30 


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31 


Effect  of  Exposure  to  Atmospheric  Air  on  the  Total  Nitrogen  Content 
of  the  Celomic  Fluid. 

In  order  to  determine  if  the  increase  in  the  carbon  dioxide  con- 
tent of  molluscan  celomie  fluid  brought  about  l)v  exposure  to 
atmospheric  air  was  in  any  way  due  to  an  increase  in  its  protein 
content  the  total  nitrogen  was  determined  in  from  25  to  100  cc 
of  composite  samples  of  celomic  fluid  taken  from  several  speci- 
mens. The  estimation  was  made  by  the  usual  Kjeldahl  method 
The  results  are  expressed  in  Table  III.     As  there  was  a  slight 


TABLE  III. 


No. 
uaed. 


1 
2 
3 

15 
8 

23 
8 
6 
6 

14 

11 
1 


Specimen. 


Total  nitro- 

Ken  prr  1(10 

cc.  of  fresh 

material. 


Schizothoerus  nuttalli. 


Saxidomus  gigantea 

Paphia  staminea 

"  u 

Cardium  corbia 

Mya  arenaria 

Polynices  lewisii  (fluid  from  foot). 


my. 

37  5 
34  0 

50.3 

70.0 

37,5 

33  6 


Total  nitro- 
gen per  100 

cc.  after 
exposure  to 

atmos- 
pheric air, 


mg. 

34.4 

51.8 

61.6 

50.4 

40  7 
9.3 


Time  in 
air. 


hrs. 

0 

0 
32 

0 
30 

0 
28 

0 
30 

0 
28 
52 


mcrease  m  two  species,  Cardium  corbis  and  Mya  arenaria,  prac- 
tically no  change  in  Saxidomus  gigantea,  and  a  slight  decrease  in 
Schtzothoerus  nuttalli  and  Paphia  staminea,  it  is  very  unlikely 
that  protein  plays  any  appreciable  part  in  the  increase  in  the 
combmed  carbon  dioxide  or  alkali  reserve  of  the  celomie  fluid  of 
the  mollusk  which  has  been  exposed  to  atmospheric  air. 

Effect  of  Exposure  to  Atmospheric  Air  upon  the  Calcium  and 
Magnesium  Content  of  the  Celomic  Fluid. 

Calcium  and  in  a  few  instances  magnesium  were  determined 
in  composite  samples  of  celomic  fluid  taken  from  several  speci- 
mens which  had  been  exposed  to  atmospheric  air  for  varying 


32 


Molluscan  C?lomic  Fluid 


S 


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J.  B.  Collip 


33 

periods  of  time.  The  method  of  McCn.ddon  (4)  was  followed, 
the  calcum  bemg  estimated  l,y  the  titration  of  the  oxalate  with 
0.1  N  potassiimi  permunpanate.     25  cc.  of  celomic  fluid   were 

wnt/l  '""^V"''r"r •  .  ^^''  ^'"'  evaporated  to  dryness  on  the 
vu^ter  bath  fused  dissolve.  1  hy  tho  aid  of  concentrated  hvdro- 
chloric  acid,  and  the  calcium  finally  precipitated  as  the  oxalate. 
The  results  are  shown  m  Table  IV.     It  will  be  noted  that,  whereas 

mollusk,  the  calcium  increases  to  a  great  extent  and  also  the 
increase  m  this  latter  constituent  is  more  or  less  parallel  with  the 
increase  ,n  the  combined  carbon  .lioxide.  It  is  therefore  evident 
that  the  great  increase  in  the  alkali  reserve  of  the  blood  of  mol- 
uscan  forms  when  expose,!  to  atmospheric  air  is  due  toan  increase 
m  the  concentration  of  bicarbonate  which  is  balanced  for  the 
\fvlT-V?' '""  '""■''^''  '"  ^^^  concentration  of  the  calcium  ions. 

oxiri/l^r/''rTi^"?"^  •'^'  '"^-  °f  ^^'"""^  ^^alculated  as 
oxide  in  the  blood  of  Saxtdomus  nuttalli  and  107  mg.  in  ^chho- 
ihoerus  nuttalli.  He  has  also  commented  on  the  very  high  I 
cium  content  of  moUuscan  blood.  I  have  failed  to  fin,l  that  the 
calcium  content  of  the  celomic  fluid  of  fresh  molluscan  forms 
diflfei^  materially  from  that  of  sea  water.  It  is  onlv  after  expc^ 
sure  to  air  that  the  calcium  content  becomes  high.  ^ 

^■^'It/f  TT  '«  f '"^•^/''i-'-'^-  ^'>  ^Pon  the  Total  Alkalinity 
and  the  Buffer  T  aluc  or  Reactivity  of  the  Celomic  Fluid. 

Lacking  the  means  of  determining  the  hydrogen  ion  concen- 
tration, a  most  important  factor  in  these  experiments  a  met  .1 
|vas  employed  to  determine  approximately  the  total  alkalinitv  o" 
tne  blood  ami  also  its  buffer  value.  The  method  adopted  was 
^nuar  to  t^at  previously  described  (1)  based  on  the  princ  " 
made  use  of  in  the  method  of  double  titration  for  bica  bona  el 

P>  n  nould  of  course  introduce  an  error  but  as  has  been  shown 
t.  .utem  content  of  the  celomic  fluid  ,loes  not  varv  to  any 
app.ociabIe  extent  an.l  therefore  approximately  the  same  de'ree 
of  error  w-ould  exist  in  all  the  titrations.  The  alkalinity  was 
determine.!  by  noting  the  amount  of  0.01  x  alkali  requ  e.  to 
produce  a  just  noticeable  pink  tint  when  phenolphthalcin  had 


THE  JOrnN-AI,  OF  BIOLOUICAL  CHEMi^ihY,    V 


Ol,   XLV,   NO.   1 


34 


Molluscan  Celoroic  Fluid 


been  added  to  the  celoniic  fluid.  The  reactivitj-  of  Moore  and 
Wilson  (7)  or  the  buffer  value  of  Sdrenson  (8)  was  determined 
l)y  titrating  from  the  phonolphthaloin  to  the  methyl  orange  point 
using  w.i,.  sulfuric  acid.  0.2  x  acid  w.-is  used  in  those  instances 
where  the  reactivity  was  of  large  proportion.  The  results  are 
shown  in  Table  IV.  It  will  be  noted  that  the  rate  of  increase  in 
the  reactivity  of  the  celoniic  fluid  is  in  close  agreement  with  the 
rate  of  increase  of  calcium  and  also  of  the  comlnned  carbon  dioxide 
content  of  the  same. 


Effect  of  Exposure  to  Air  Followed  by  Submersion  in  Fresh  Water. 

Table  \  illustrates  that,  whereas  exposure  to  air  causes  a  rapid 
increase  in  the  alkali  reserve  of  the  celoniic  fluid,  the  subsequent 
immersion  in  fresh  sea  water  causes  a  return  to  approximately 
the  normal  value  for  this  factor. 

TABLE  V. 


Specimen. 

COi  content  of 
lOOec.of  fluid 
equilibrated 
with  atmos- 
pheric air. 

Remarks. 

Mya  arenaria. . 

U                   it 
«                      X 

ii              t( 
<(             « 

cc. 

8.2 
32.0 
23.3 
14  9 

9.2 

Fresh. 

72  hrs.  in  glass  container  in  laboratory. 

Submerged    4  hrs.  in  fresh  sea  water. 

a                  ^Q      i(       ((       a           It         it 
11                 nj      tt       It       tt           it        it 

Effect  of  Srtbmrrsion  in  Sea  Water  in  a  Sealed  Container. 

Several  fresh  specimens  of  Mya  arenaria  were  immersed  in  a 
relatively  small  volume  of  sea  water  in  a  cylindrical  glass  con- 
tainer which  was  then  tightly  sealed.  After  varying  periods  of 
time  the  celomic  flui<l  of  the  specimens  was  examined.  An 
analysis  of  the  .«ea  water  vhich  was  used  in  the  experiment  was 
also  made.  The  results  are  expre.>^.sed  in  Table  VI.  A  few 
experunents  were  carried  out  in  which  lioiled  out  soa  water  was 
used  in  iilace  of  fresh  sea  water.  In  on'  instance  sea  water,  the 
buffer  value  of  which  had  been  greatly  increased  l)y  the  addition 
of  5  gm.  of  l)asie  sodium  i)hosphate  per  liter,  was  used.     It  will 


J.  B.  CoUip  35 

be  noted  that  the  J«>havior  of  Mya  arenarm  kept  in  a  scaled  con- 
taincr  in  either  boiled  or  fresh  sea  water  is  ver^-  similar  to  that 
ol.ser%ed  when  the  specimens  were  kept  in  the  air.    The  alkali 
resen-e  and  the  calcium   content  of  the  celomic  fluid  mount 
steadily  until  the  animal  dies.    The  increase  in  the  carbon  diox- 
ide and  calcium  content  of  sea  water  is  also  considerable.    The- 
values  for  these  two  factors  in  sea  water  are,  however,  lower  in  the 
case  of  the  celomic  fluid  except  after  long  submersion  of  the 
specimens  in  which  instance  there  is  a  tendency  for  the  concen- 
trations of  these  latter  substances  in  the  celomic  fluid  and  sea 
water  to  equalize.    The  addition  of  basic  sodium  phosphate  to 
the  boiled  sea  water  used  in  one  experiment  did  not  materially 
alter  the  results.    A  dense  precipitate  was  formed  in  the  sea 
water  in  this  experiment  which  consisted  for  the  most  part  of 
calcium  phosphate. 

If  one  considers  the  increase  in  the  combined  carbon  dioxide  in 
the  sea  water  used  in  an  experiment  one  finds  that  there  is  very 
little  difference  in  the  rate  of  increase  in  this  factor  in  specimens 
exposed  to  air  and  in  specimens  submerged  in  a  relatively  small 
yolmne  of  sea  water.  Thus  in  Experiment  I.  Table  Vl  an 
increase  of  29.3  volumes  per  cent  was  noted  in  the  combined 
carbon  diox.de  content  of  the  celomic  fluid,  while  an  increase  of 
fuJly  20  volumes  per  cent  took  place  in  the  sea  water.  The 
total  bulk  of  the  eight  specimens  used  in  this  experiment  was 
550  cc.  while  the  .volume  of  sea  water  used  was  750  cc  There 
was  therefore  an  increase  of  150  cc.  of  combined  carbon  dioxide 
due  to  the  actiyity  of  the  specimens  over  and  above  the  increase 

d,  *nt  '°,  «-'''      ?"^'  ^"^'^^    ^^^  ^^^"«  «f  *he  eight  specimens 
displaced  8o  cc.  of  water.    Not  allowing  for  the  water  entrapped 
in  the  mantle  cavities,  there  were  465  cc.  of  clam  tissue  present 
m  this  experunent  so  that  the  increase  of  150  cc.  of  carbon  dioxide 
found  m  the  sea  water  would  mean  that  32  cc.  of  this  carbon 
dioxide  had  resulted  from  the  activity  of  each  100  cc.  of  clam 
tissue.    This  added  to  the  carbon  dioxide  content  of  100  cc   of 
celomic  fluid  indicates  that  approximately  01.3  cc.  of  combined 
carbon  dioxide  resulted  from  the  aclivitv  for  40  hours  of  100  cc 
of  clam  tissue,  an  amount  which  is  in  close  agreement  with  the 
obsen;ed  increase  of  the  volume  per  cent  of  carbon  dioxide  in  the 
celomic  fluid  of  similar  specimens  exposed  to  atmospheric  air  in 
a  closed  vessel  for  a  corresponding  period  of  time  (Table  IV) 


36 


Molluscan  Celomic  Fluid 


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J.  B.  CoUip 


37 


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38 


MolliLscan  Celomic  Fluid 


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J.  B.  follip 


39 


KJTirl  of  Suhmimiun  in  l)ixtilli<f  Wnlir. 

The  rosiilfM  of  soruo  cxprrinjcnts  in  which  HjM'(irii«'ii,M  oi    \fya 
urcmrin  woro  hiiimierged  in  fresh  (li.still«i  water  in  a  rUk*  con- 
tainer and  kept  at  the  temperature  «if  surface  sea  water  ap/war 
in  Table  VII.     The  conihined  eMrLon  dioxide  and  ealciiini  f^n- 
tent  of  celoniie  fluid  rise  after  nnieh  tlie  xanie  niaritier  aH    vas 
oh8er\-ed  when  specimens  were  placed  in  .-ither  frexh  or  Unm\ 
sea  water.     Kxperiments  I  and  II,  Tal.lc  VII,  are  of  intercHt  in 
that  they  show  that  a  fall  had  taken  place  in  the  concentration 
of  magnesium  in  the  celoniie  fluid  of  the  specimens,  while  a  riw 
occurred  in  the  concentration  of  j-alcium.     There  i    •.!.,»  a  niarkcH 
difTen-nce  in  the  total  alkidinity  and  in  the  read 
and  the  celoniie  fluid.     The  chlorine  content  ol 
and  of  the  water  in  which  the  specimens  of  .1/ 
suhmerKcd  was  determined  in   Kx|)erimeiits   II 
VII.     The  manner  in  which  the  concentratioi, 
the  celoniie  fluid  is  kept  :it  a  relatively  liifth  le\ 
eumstnnces  ohtaining  in    iiose  experiments  is  ion 
(9)  has  shown  that  the  addtn-for  muscle  of  tlK- 
nmria  is  jM-culiarly  resistant  to  hypertonic  ,^4»atir«,s  ,rf  ^,J,Iiw, 
chloride  and  to  (knihie  strength  sea  water,  flie  cnrrnti     m^ 
sodiun»  chloride  -:  Ini;  to  only  ahout  one-half   that  ,.f  ti.^  ^„, 
rounding  mediuia  >!so  found   that  the     ..antic  i:.    it....^., 

imiK-rmeal.le  to  .sodiu,,  .oride.  The  al.ilin  ^^(  M ,,  ,.,,/« 
to  withstand  immersion  iii  water,  the  osinoii,  „n-j^niv  •  Jieh 
is  verj'  low,  is  of  interest  in  the  lijjht  of  th.  na' On  ^ 

Effect    of    Kj-posiiir    to    (I    llii<lro(i<       Alitios/,i„ 

Several  specimens  of  .1///,/  anmnin  were  kept  in  a  MM:.^  ue 

of  .sea  water  for  2  hour-  in  order  that  the  ow^en  .-..nt.ni  ..jr 

tissues  should  he  redm,  1  to  a  low  level.  ( h.e  .specim.  i  .  ifien 
hied  as  a  control,  tlu  -thers  were  placed  in  a  hvdron.  atmo.s- 
sphere  over  alkaline  pyiofrallic  acid,  tw..  separate  containers 
hemg  u.sed;  One  uroup  of  three  was  exposed  for  2(>  lioiir^-  the 
other  for  48  hours.  The  results  of  the  analy.s(.s  of  the  .vl'omic 
(huds  of  these  specimens  are  shown  in  Tal.le  VIII.  '  Tl..  maxi- 
mum and  minimum  temjieratures  for  the  2  days  durinfi  which 


the  wat*^ 
•iniic  fluid 
**iri(i  were 
'fl,  'I  iihie 
hlori(i  in 
the     ir- 


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40 


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41 


thi.H  experiment  wjim  jwrfoniKHl  were  on  the  lut  day  24.4»  and 
lO.ft'C,  on  the  2n.|  duy  2a.9'  und  11.4"f'.  It  will  l.e  noticed 
that  the  incnmse  ohwem-d  in  the  concentration  of  l)icttrl)onate  in 
the  cclomie  fluid  is  very  nuirketl.  It  iM  not,  however,  so  Rreat 
as  that  which  is  found  when  tlie  siH'ciniens  areexpose«l  toatn»o«- 
piieric  air.  The  increase  in  tlie  calciuni  and  magnesium  is  com- 
parable to  that  ol)8er\-cd  for  bicarbonate. 

Effect  of  Exposure  to  a  Nitrogen  Atmosphere. 

Three  Bpecimena  of  Mya  arenaria  were  placed  in  a  small 
desiccator  "ontaining  a  concentrated  solution  of  pyrogallic  acid 
in  40  i)er  cent  sotlium  hydroxide.    A  glass  tute  was  so  attached 


TABLE  IX. 

No. 
iMed. 

Sp«cimrn. 

COiper 

lOOcc.of 
tiuid  <><iuili- 
brstnl  with 

■tmiM- 
pheric  air. 

Remark  •. 

3 
3 

3 

Mya  arenaria 
•<           II 

II           II 

ee. 

SO 
34  8 
17.8 

Fresh  control. 

25  hrs.  in  glass  container  in  laboratory 
25    "     "  desiccator  over  alkaline  pyro 
gallol. 

to  the  exhaust  cock  that  as  oxygen  was  ab.sorl)ed  the  air  which 
enten'd  first  bubbled  through  the  alkaline  pyrogallol  in  the  bottom 
of  the  desiccator.  Three  other  s|)eciinens  of  like  size  were  placed 
at  the  same  time  in  a  gla-ss  container  which  was  kept  at  the  same 
temiierature  as  the  desiccator  for  the  duration  of  the  experiment. 
After  25  hours  the  specimens  were  bled  and  an  analysis  was 
made  of  the  composite  .samples  of  celoniic  fluid.  The  results 
are  expre.s.sed  in  Table  IX.  It  will  be  observe<l  that  the  com- 
bined carbon  dioxide  of  the  celomic  fluid  did  not  increase  to 
the  same  extent  in  the  sjiecimens  wi.icli  wore  kept  in  the  desic- 
cator over  alkaline  pyrogallol  as  it  did  in  the  controls  which  were 
kept  in  the  air. 


42 


MoUuscan  Celomic  Fluid 


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J.  B.  Collip 


43 


Comparison  of  the  Carbon  Dioxide  Content  of  Cclomic  Fluid  and 
Other  Fliiidfi  Obtained  from  Mollusca. 

A  comparison  was  made  between  the  carbon  dioxide  content 
of  the  celoinic  fluid  and  other  fluids  of  ilifforcut  iloUusca.  The 
rcsuhs  of  this  study  are  shown  in  Tabic;  X.  It  was  found  that 
the  carbon  dioxide  content  .f  the  fluid  which  exuded  from  the 
exhalant  siphon  of  fresh  Mi/a  arenaria  was  just  sUghtly  higher 
tlian  that  of  sea  water.  >\lien  specimens  of  this  species  were 
exposed  to  atmospheric  air  for  some  time  the  l)icarl)onate  content 
in  the  fluid  of  the  mantle  cavity  closely  approximated  that  in  the 
•■elomic  fluid.  Somewhat  similar  observations  were  made  on 
the  auiphincuran  form  Crijptochiton.  The  fluid  in  the  foot  of 
the  large  gastropod  Pohjnicen  leirisii  l)ears  somewhat  different 
relation  to  the  blood  and  celomic  fluid  of  this  form  as  far  as  the 
bicarbonate  content  is  concerned  from  that  which  holds  in  the 
fluid  between  the  mantle  cavity  and  the  blood  and  celomic  fluid 
of  a  pelecyi)od  such  as  Mija  arenaria.  The  combined  carbon 
dioxide  content  of  the  fluid  of  the  foot  of  Poli/nices  leirixii  approx- 
imates the  value  obs(>r\-ed  for  tlu;  celomic  fluid.  The  drawing 
in,  therefore,  of  the  foot  in  this  form  causes  a  considerable  decrease 
in  the  total  alkali  reserve  of  the  animal.  It  is  of  interest  in  this 
connection  to  note  that  this  animal  does  not  withdraw  its  foot 
imless  subjected  to  rather  violent  irritation. 


Effect  of  Submersion  of  Dead  Specimenx  of  Mya  arenaria  in  Sra 

Water. 

The  results  of  two  experiments  are  shown  in  Table  XI.  It  is 
evident  that  the  bicarbonate  content  of  sea  water  in  which  ilead 
clams  are  immersed  rises  quite  rapidly  once  decomposition  has 
set  in.  It  will  be  noted,  however,  that  there  is  little  change 
during  the  first  21  hours  submersion.  The  behavior  of  pelecypod 
mollusks  exposed  to  air  or  submerged  in  a  relatively  small  vol- 
mne  of  water  is  therefore  quite  distinct  from  that  of  dead  clams 
which  are  undergoing  decomposition. 


44 


AloUuscan  Celomic  F.  lid 


TABLE  XI. 


Specimen. 

Total  COi 
per  100  re. 

Remarks. 

Experiment  I. 
4  Mya  arenaria  (175  cc.) 

Sea  water  (boiled) 

cc. 

3  3 
4.7 

42.0 

55.0 
07.0 
72.0 

2.8 

3  3 

11  7 

35.0 

Placed  in  275  cc.  of  boiling  sea 
water. 

«<       i(            11 

After  24  hrs.  No  decomposi- 
tion of  clam  tissue. 

After  48  hrs.  Decomposition 
clearly  manifested. 

After  72  hrs 

i(            u                   u 

«           tl                  it 

It         tt               tt 

"        96     " 

tt         tt               tt 

"     144    " 

Experiment  II. 
10  Mya  arer"  ia  (500  cc.) 

Sea  water  (boiled) 

Placed   in  700  cc.  of  boiled  sea 
water  containing  9.5  per  cent 
alcohol. 

U             It                     tt 

After  24  hrs 

l<        11              It 

"     48    "       .Milky,  decompo- 
sition evident. 
After  96  hrs 

«        (1              tt 

DISCUSSION. 

As  has  already  l)cen  indicated  the  marked  increase  in  the 
l)icarl)onate  content  of  the  celomic  fluid,  and  therefore  in  all 
probal)ility  of  the  hlood  and  tissues  of  the  calcareous  shelled 
pelecypod  moUusks  and  the  arthropod  Balnnm  aquilla  on  expos- 
ure to  air  is  quite  opposite  to  the  effect  ohserv'ed  in  fishes  when 
the}'  are  removed  from  their  natural  habitat.  This  phenomenon 
is  undoubtedly  associated  with  the  presence  of  a  calcareous  shell 
the  calcium  carbonate  of  which  furnishes  an  alkali  reserve  which 
is  added  to  that  of  the  body  fluids  and  tissues,  and  which  it 
appears  can  be  readily  utilized. 

As  specimens  of  Myn  arrnnrin  appear  to  remain  practically 
normal  even  after  long  exposure  to  atmospheric  air  there  is  no 
reason  to  supjwse  that  any  material  change  has  been  effected  in 
their  metabolic  proce.s-ses  as  a  result  of  the  change  in  environment. 
If  one  assumes,  therefore,  that  combustion  still  i)roceeds  in  the 


^■^ft:.. 


J.  B.  CoUip 


45 


exposed  specimens,  then  the  increase  in  the  bicarbonate  content 
of  the  body  fluids  can  be  explained  according  to  the  equation 

CO2  +  H2O  +  CaCO,  t=;  Ca(HC05): 

The  carbon  dioxide  resulting  from  the  respiratory  process 
would,  l)y  shjihtly  increasing  the  hydrogen  ion  concentration, 
dissolve  calcium  carbonate  from  the  shell  and  the  concentration 
of  the  calcium  ions  and  of  i)icarbonato  ions  would  therefore 
steadily  rise  as  combustion  in  the  tissues  proceeded.  The  amount 
of  carbon  dioxide  actually  excreted  from  the  specimens  in  the 
gaseous  form  was  not  determined.  If  this  factor  were  known 
one  could  calculate  the  intensity  of  metaljolism  in  these  forms 
by  considering  the  amount  of  carbon  dioxide  excreted  in  addition 
to  the  amount  retained  as  l)icarbonate.  It  would  appear  that 
50  per  cent  of  the  increase  observed  in  the  carbon  dioxide  content 
of  the  celomic  fluid  is  due  to  carbon  dioxide  formed  by  combus- 
tion in  the  tissues,  while  the  remaining  50  per  cent  results  from 
the  solution  of  calcium  carlionate  of  the  shell. 

The  degree  of  alkalinity  of  the  celomic  fluid  determined  by 
titration  is  l)y  no  means  an  indication  of  the  hydrogen  ion  con- 
centration, but  the  ratios  observed  between  the  alkalinity  figures 
and  the  reactivity  values  suggest  that  no  marked  increase  in  the 
hydrogen  ion  concentration  takes  place  during  the  early  part  of 
the  exposure  at  least.  The  reactivity  or  buffer  value  is,  in  nearly 
every  instance,  in  close  agreement  with  the  calcium  content  and 
the  carbon  dioxide  concentration  of  the  celomic  fluid. 

The  increase  in  the  alkali  reserve  as  indicated  by  an  increase 
in  bicarbonate  concentration  in  specimens  exposetl  to  atmospheric 
air  is  due  for  the  most  i)art  to  increase  in  the  calcium  content. 
Magnesium,  which  in  the  normal  animal  in  its  natural  habitat 
exceeds  calcium  in  the  degree  of  its  concentration,  and  therefore 
balances  a  greater  proportion  of  bicarbonate  ions  than  does 
calcium,  increases  only  slightly  as  comparei!  with  calcium  when  a 
specimen  is  exposed  to  air.  It  is  prol)al)le  that  the  relative 
increase  in  calcium  and  magnesium  concentrations  under  these 
circumstances  is  somewhat  similar  to  the  relative  amounts  of 
these  substances  in  the  shell  from  which  solution  of  bicarbonate 
is  taking  place.     It  is  of  interest  to  note  here  that  no  increase 


46 


Molluscan  Celomic  Fluid 


was  ohsorvcd  in  the  concentration  of  n.aRnosiuin  in  the  cockle 
(tardiiini  corhis)  on  exposure  to  air. 

As  specimens  suhnierRed  in  boiled  sea  water  and  kept  in  a 
sealed  container  cntinne  to  develop  an  increased  carbon  dioxide 
content,  calcunn  concer.t ration,  and  buffer  value,  after  much 
the  same  manner  as  specimens  exposed  to  atmospheric  air,  and 
since  a  similar  effect  is  manifeste.1  by  specimens  kept  in  an  atmos- 
phere  of  hydrosen  or  nitrogen,  one  is  led  to  ask  the  question  "Can 
anaeroluc  respiration  be  manifested  l>v  these  forms?" 

If  one  considers  the  results  of  an  experiment  recorded  in  Table 
Mil,  one  fimls  that  after  48  hours  in  a  hydrogen  atmosphere 
the  combined  cari)on  dioxide  ro.sc  from  8.2  to  45.5  volumes  per 
cent,  or  an  observed  increase  of  37.3  volumes  per  cent.     If  one 
a.ssumes  that  aerobic  respiration  was  taking  place  and  that  car- 
bohyilrates  were  I,eing  burned,  then  a  volume  of  oxvgen  equiva- 
lent to  the  volume  of  carbon  dioxide  produced  would  l,e  required. 
It  ;)0  per  cent  of  the  obser^-ed  increa.se  in  the  combined  concen- 
tration of  carbon  ilioxide  is  indicative  of  the  amount  of  this  sub- 
stance  prmluced  due  to  combustion  then  an  amount  of  oxygen 
equivalent  to  .",0  ,,of  cent  of  37.3  volumes  per  cent,  or  18  05 
volmnes  per  cent,  would  be  required.     As  the  specimens  used 
in  this  e.xijerunent  were  kept  in  a  small  volume  of  sea  water  for 
2  hours  before  they  were  transferred  to  a  hydrogen  atmosphere, 
one  fails  to  see  how  any  appreciable  amount  of  oxvgen  covld  be 
contamed  m  the  tissues  of  the  specimens.    As  there  is  no  apparent 
source  lor  18.05  volumes  per  cent  of  oxygen  in  these  clams  it  is 
therefore  evident  that  thej-  must  be  respiring  anaerobically  or 
ese  the  increase  in  the  carbon  dioxide,  calcium,  and  buffer  value 
of  the  celomic  fluid  is  due  to  some  other  cause  than  that  suggested 
ear  her  in  the  paper.     Decomposition  of  the  clam  tissue  can  be 
exclu,  ed  smee  the  specimens  were  very  active,  resijonding  to 
s  inmlation  like  normal  animals,  after  they  were  removed  from 
the  hydrogen  atmos[)here. 

There  is  the  possibility  of  (he  solution  of  the  calcium  carbonate 
ot  the  shell  due  simply  to  the  .solvent  action  of  the  tissue  fluids 
containing  free  carbon  dioxide.  This  would  result  in  the  forma- 
tion of  calcumi  carbonate  the  solubility  „f  which  is  considerablv 
increased  by  an  excess  of  carbon  dio.xide  in  the  water  (10)  In 
dealing  with  a  closed  system,  however,  s.ich  as  the  individual 


•  '--.m 


J.  B.  Collip 


47 


clam  in  an  atmospliprc  of  hydroKon,  tlio  solution  of  calcium  car- 
bonate due  to  the  solvent  action  of  the  free  carbon  flioxidc  would 
require  a  constant  supply  of  the  latter  if  the  process  is  to  con- 
tinue; otherwise  etiuilibrum  would  be  established  between  the 
dissolved  bicarbonate,  the  calcium  carbonate  of  the  shell,  and 
the  free  carl  .on  dio>dde,  and  no  further  increase  would  be  mani- 
fested unless  this  balance  were  disturbed.  It  will  be  noted  that 
the  rate  of  increase  in  the  carbon  dioxide  content  mid  the  calcium 
concentration  of  the  blood  is  for  a  considerable  period  practically 
constant.  If  respiratory  activity  account.s  for  the  increase  in 
bicarbonate  concentration  of  the  tissue  fluids,  then  this  uniform- 
ity in  the  rate  obseri-ed  would  fit  in  well  with  the  fairly  constant 
rate  of  metabolism  which  might  be  expected  under  such  circum- 
stances, anaerobic  respiration  being  possible. 

The  fact  that  the  rate  of  increase  in  the  bicarlfonate  concen- 
tration, the  calcium  content,  and  the  buffer  value  is  greater  in 
air  than  it  is  in  either  hydrogen  or  nitrogen  would  indicate  that 
absence  of  oxygen  does  exert  an  influence  on  the  intensity  of  the 
metai)olic  processes  but  by  no  means  causes  a  complete  cessation 
in  the  respiratory  function. 

The  extreme  sensitivity  of  the  cockle  and  the  horse  clam  to 
expo.sure  to  air  at  the  prevailing  summer  temperatures  made 
experiments  with  these  forms  of  the  same  type  as  were  conducted 
with  Myn  arcnaria  temporarily  impossible.  Neither  of  these 
forms  is  normally  submitted  to  the  same  degree  of  low  oxygen 
tension  as  is  Mija  airnaria.  It  is  hoped  that  experimental  work 
of  a  similar  nature  to  that  carried  out  with  Mya  aremria  may  be 
done  on  other  forms  at  a  more  favorable  time  of  the  year. 

It  has  long  been  known  that  animals  show  a  very  unequal 
resistance  to  lack  of  air.  liunge  (11)  in  his  work  upon  respira- 
tion of  intestinal  parasites  and  nmd-dwelling  organisms  showed 
that  parasites  in  the  intestine  of  warm  i)looded  animals  must 
live  practically  in  the  absence  of  o.xygen,  while  wonns  living  in 
the  nmd  were  also  subject  to  similar  conditions,  decomiwsition 
processes,  with  the  formation  of  reducing  substances,  keeping 
the  oxygen  absent.  Packard  (12)  found  that  worms  and  mud- 
dwelling  Crustacea  are  resistant  to  the  lack  of  o.xygen  for  some 
time. 


48 


MoUuscan  Celomic  Fluid 


The  ability  of  an  animal  to  resist  a  lack  of  oxygen  may  or 
may  not  he  connected  with  an  anaerobic  respirator^'  mechanism. 
If  one  finds  complete  evidence  of  metabolism  in  an  animal  exposed 
to  anaerobic  conditions,  then  anaerobic  resjuration  woidd  be 
indicated.  Such  seems  to  be  the  condition  in  the  ca.se  of  Mya 
arffiario.  Crustacean  types  which  were  exposeil  to  the  air  or 
submerged  in  boiled  sea  water  died  within  a  few  hours.  The 
carbon  dioxide  content  of  the  blood  was,  however,  practically 
unaltered  by  such  procedures.  These  forms  do  not  use  the 
calcium  carbonate  of  their  carapace  as  a  protective  measure  when 
removed  from  their  normal  habitat. 

In  the  light  of  the  results  of  the  experiments  which  have  so 
far  been  conducted  upon  Myp  armaria  the  writer  has  tentatively 
to  conclude  that  indiviihials  of  this  species  behave  as  facultative 
anaerobic  organisms.  It  is  realized,  however,  that  in  these  pre- 
liminary experiments  absolutely  anaerobic  conditions  were  not 
secured.  It  is  the  intention  to  continue  this  work  at  another 
time  when  it  is  hoped  to  <letcrmine  the  hydrogen  ion  concentra- 
tion of  the  celomic  fluid  and  the  rate  of  oxygen  consumption 
under  various  conditions. 


SUMMARY. 

1.  Calcareous  shelled  pelecypod  Mollusca  and  the  arthropod 
Balanus  aquilla  have  in  the  calcium  carbonate  of  their  shells  a 
potentially  great  alkali  reserve. 

2.  I'Aposure  of  these  forms  to  atmospheric  air  causes  a  marked 
increase  in  the  combined  carbon  dioxide  of  the  celomic  fluid. 

3.  There  is  imder  these  circumstances  a  parallel  increase  in  the 
calcium  concentration  and  the  buffer  value  of  the  celomic  fluid. 

4.  Various  other  marine  forms  studied  did  not  so  react. 

").  There  is  no  increase  in  the  total  nitrogen  of  the  celomic  fluid 
of  the  pelecyiiod  Mollusca  exposed  to  atmospheric  air. 

G.  Mya  arenaria  is  particularly  resistant  to  long  exposure  to 
atmospheric  air. 

7.  When  specimens  of  Mya  arenaria  are  placed  in  a  relatively 
small  volume  of  fresh  sea  water,  boiled  sea  water,  distilled  water, 
or  in  a  hydrogen  or  a  nitrogen  atmospliere  much  the  same  reac- 
tion is  observed  as  when  specimens  are  exjiosed  to  atmospheric 
air. 


J.  B.  CoUip 


49 


8.  The  rate  of  increase  in  the  content  of  carbon  dioxide,  the 
calcium  concentration,  and  the  l)uflfer  value  of  the  celomic  fluid 
under  all  the  above  conditions  is,  during  the  first  period  of  sev- 
eral hours,  constant. 

9.  The  rate  of  increase  in  the  rate  of  combined  carbon  dioxide, 
the  concentration  of  calcium,  and  the  buffer  value  is  not  so  great 
in  a  hydrogen  or  nitrogen  atmosphere  as  it  is  in  air. 

10.  It  is  suggested  that  Myn  armaria  is  a  Tacultative  anaerobic 
organism  which  continues  to  produce  carbon  dioxide  under 
anaerobic  conditions. 

In  conclusion  I  wish  to  exiiross  my  thanks  to  Dr.  C.  McLean 
Fraser,  the  Curator  of  the  Biological  Station,  Departure  Bay, 
British  Columbia,  for  his  cooperation  during  the  carrying  out  of 
the  experimental  work  reportc'  in  this  communication. 

My  thanks  are  also  due  to  the  Biological  Board  of  Canada, 
by  whom  the  exjicnses  in  connection  with  this  investigation  1  e 
been  dcf raved. 


BIBLIOGRAPHY. 

1.  Collip,  J.  B..  J.  Biol.  Chem.,  1920,  xliv,  329. 

2.  Van  Slyke,  D.  D.,  J.  Biol.  Chem.,  1917.  xxx,  347. 

3.  Van  Slyke,  D.  D.,  and  CuUen,  G   E.,  J.  Biol.  Chem.,  1917,  xxx,  289. 

4.  McCrudden,  F.  H.,  J.  Biol.  Chem.,  1911-12.  x,  187. 

5.  Myers,  R.  G.,  J.  Biol.  Chem.,  1920,  xli,  119. 

6.  Brown,  H.  T.,  and  I']8combe,  F.,  Phil.  Trans.  Roii.  Soc,  1900,  cxciii, 

289. 

7.  Moore,  B.,  and  Wilson,  F.  P.,  Biochem.  J.,  1906,  i,  207. 

8.  Sorensen,  S.  P.  L.,  in  Moore,  B.,  Prideaux,  E.,  and  Herdman.  G., 

Trans.  Biol.  Soc.  Liverpool,  1915,  xxix,  174. 

9.  Meigs,  E.  B.,  Am.  J.  Physiol.,  1914,  xxxiii.  p.  xxii. 
10.  Reuter,  M.,  Z.  anorg.  Chem.,  1898,  xvii,  170. 

11;  Bunge,G.,Z./)A.vsiV»/.C/icm.,lS83-t^4.viii,48:188S.xii,565;1890,xiv,  318. 
12.  Packard,  \V.  H.,  .4m.  J.  Physiol.,  1905-06,  xv,  30. 


I 


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